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Journal of Bacteriology, January 2003, p. 496-503, Vol. 185, No. 2
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.2.496-503.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of a Novel Intracellular Endopeptidase of the {alpha}/ß Hydrolase Family from Streptomyces coelicolor A3(2)

István Nagy,1,2 Tisha Banerjee,1 Tomohiro Tamura,2,{dagger} Geert Schoofs,1 Ann Gils,3 Paul Proost,4 Noriko Tamura,2,{dagger} Wolfgang Baumeister,2 and René De Mot1*

Centre of Microbial and Plant Genetics,1 Laboratory for Pharmaceutical Biology and Phytopharmacology,3 Laboratory of Molecular Immunology, Rega Institute for Medical Research, Catholic University of Leuven, Leuven, Belgium,4 Department of Molecular Structural Biology, Max-Planck-Institute of Biochemistry, Martinsried, Germany2

Received 3 June 2002/ Accepted 21 October 2002

In a proteasome-lacking mutant of Streptomyces coelicolor A3(2), an intracellular enzyme with chymotrypsin-like activity, absent from the wild type, was detected. Complementation that restored proteasome function did not suppress expression of the endopeptidase. Since the enzyme was not found in two other S. coelicolor proteasome mutants, its expression probably resulted from a secondary mutation arisen in the proteasome mutant. Purification of the endopeptidase revealed its identity to SCO7095, a putative hydrolase encoded by the S. coelicolor A3(2) genome with no known homologue. Based on the prediction of a Ser-Asp-His catalytic triad and an {alpha}/ß hydrolase fold, SCO7095 was assigned to peptidase clan SC. N-terminally His-tagged SCO7095 was efficiently expressed in Escherichia coli cells and purified for further characterization. Although SCO7095 is distantly related to several proline iminopeptidases, including Thermoplasma acidophilum tricorn-interacting F1, no aminopeptidase activity was detected. On synthetic substrates, the monomeric enzyme exhibited not only chymotrypsin-like activity but also thrombin-like activity.

* Corresponding author. Mailing address: Centre of Microbial and Plant Genetics, Catholic University of Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee, Belgium. Phone: 32 16 32 9681. Fax: 32 16 32 1963. E-mail: rene.demot{at}agr.kuleuven.ac.be.

{dagger} Present address: National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, Japan.

Journal of Bacteriology, January 2003, p. 496-503, Vol. 185, No. 2
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.2.496-503.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hong, B., Wang, L., Lammertyn, E., Geukens, N., Van Mellaert, L., Li, Y., Anne, J. (2005). Inactivation of the 20S proteasome in Streptomyces lividans and its influence on the production of heterologous proteins. Microbiology 151: 3137-3145 [Abstract] [Full Text]  


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